Fig. 4

Molecular insights into BafA1-mediated metabolic rewiring of GSCs. a A metabolic gene expression panel (n = 748 genes; see also Suppl. Table S4) was analyzed in GSCs treated with vehicle (Ctrl) or 5 nM Bafa1 for 48 h. List of genes whose expression was significantly up (adj p value < 0.05 and log2FC > 0.5) in Ctrl-GSCs or in BafA1-GSCs were separately imported in Reactome web-tool and the pathway analysis was performed with voronoi pathway visualization. The most relevant activated signaling in Ctrl- or BafA1-GSCs are shown. For the complete voronoi charts please refer to Suppl. Figure 7. b Transcripts belonging to the indicated hallmark and up (red) or down-modulated (green) in BafA1-treated GSCs are shown (see also Suppl. Table S5). c The expression of the activating transcription factor 4 (ATF4) was analyzed in Ctrl- and BafA1-treated GSCs. Data are showed as violin plots and each dot is a sample. **, p = 0.002 by Mann–Whitney U test. d Ingenuity pathway analysis was performed integrating gene expression (GEX) and metabolic data. Pathway predicted as activated or inhibited (positive and negative z-score, respectively) in BafA1-GSCs are shown (see also Suppl. Table S7). f Protein de novo synthesis was evaluated in GSCs incubated for 24 h with vehicle (Ctrl) or 5 nm BafA1 using fluorescence microscopy. The mean fluorescence intensity (MFI) was measured and quantified by Image J software (bottom panel). Scale bar, 20 µm. ***, p < 0.0001 by Mann–Whitney U test